LOCUS       V004436                 6100 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004436
VERSION     V004436
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6100)
  AUTHORS   Heap JT, Ehsaan M, Cooksley CM, Ng YK, Cartman ST, Winzer K, Minton
            NP.
  TITLE     Integration of DNA into bacterial chromosomes from plasmids without
            a counter-selection marker
  JOURNAL   Nucleic Acids Res. 40 (8), E59 (2012)
   PUBMED   22259038
REFERENCE   2  (bases 1 to 6100)
  AUTHORS   Heap JT, Ehsaan M, Cooksley CM, Cartman ST, Minton NP.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-JAN-2011) School of Molecular Medical Sciences, The
            University of Nottingham, Centre for Biomolecular Sciences,
            University Park, Nottingham, Nottinghamshire NG7 2RD, United Kingdom
REFERENCE   3  (bases 1 to 6100)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6100)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic
            Acids Res."; date: "2012"; volume: "40"; issue: "8"; pages: "E59"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (11-JAN-2011) School of Molecular Medical Sciences, The University
            of Nottingham, Centre for Biomolecular Sciences, University Park,
            Nottingham, Nottinghamshire NG7 2RD, United Kingdom"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6100
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     gene            9..546
                     /gene="pyrE fragment"
                     /label="pyrE fragment"
     misc_feature    9..308
                     /gene="pyrE fragment"
                     /note="mediates homologous recombination with the
                     chromosome of Clostridium difficile 630"
     primer_bind     complement(665..681)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     misc_feature    829..2028
                     /note="mediates homologous recombination with the
                     chromosome of Clostridium difficile 630"
     CDS             1040..1966
                     /codon_start=1
                     /gene="CD0188"
                     /product="putative ketopantoate reductase"
                     /label="CD0188"
                     /protein_id="AFA34970.1"
                     /translation="MKNQLNENNKKVVFFGVGAVGATFAEQFFNSKYDFKILCDNERKK
                     RYLEEGFIINGKRYDFDYVTKDEYKQEADFIIIGLKYNNLKENIKELDGLVGKNTVIMS
                     LLNGVDSEEIIGERFGIEKMVYSYVTNIDAKKINNNIIHTTNGIIVFGNKDNSEDRKTN
                     IITEVFDDVNIEYTLSKDIQRDMWWKYMVNIGVNQTSAILGAPYGVFQSSEHLRELAKS
                     AMREVVAIAQAKDISLTEDDVEHSLHRILEHSKEGRTSMLQDVEAHRLTEVDMFSKNIC
                     KLGKKYNIPTPINQTFFYMIKVIESRF"
     gene            1040..1966
                     /gene="CD0188"
                     /label="CD0188"
     CDS             2755..3072
                     /codon_start=1
                     /gene="repH"
                     /product="plasmid replication protein"
                     /label="repH"
                     /protein_id="AFA34971.1"
                     /translation="MKRRAYMKTCKNCKEFIKDTEICKIHSLMIHDKTVATYCSKYNES
                     RCLHKGKVQCINCSKMNRYGWCAIKMRCFTEEEQKKERTCIKYYARSFKKAHVKKSKKK
                     K"
     gene            2755..3072
                     /gene="repH"
                     /label="repH"
     CDS             3744..4364
                     /gene="catP"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from Clostridium
                     perfringens. Accession#: P26826"
     rep_origin      4544..5132
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     oriT            5453..5562
                     /label="oriT"
                     /note="incP origin of transfer"
     CDS             5595..5963
                     /label="traJ"
                     /note="oriT-recognizing protein"
ORIGIN
        1 cctgcagggg agggacattt tttattatct tcaggaaaac acagtaatag atacgtacaa
       61 tgtgcaaaag tattaagatt cccagaatat gctgctgagg tattaagtac agttgttgaa
      121 caaataaaag acttagatat agacttagta gtaggaccag ctatgggtgg agtaatagtt
      181 tcttatgagt taggaagaca attaggaaaa gaagctgtat ttactgagag aaaagacaat
      241 acaatggagt taagaagagg atttgaagtt aaaaaaggag caaagataat aattgctgaa
      301 gacgttgtaa ctactggtaa atcaactatg gagacaaaaa gagtattaga agccttaggt
      361 ggagaagttg taggtgttgc atgtatagca gatagaacta atcatgatat aggtatgcct
      421 atatatagtg ctataaaact tgatattcaa gtttatgaat ctgatgagtg tcctttatgt
      481 aaggaaggaa aattaccagt tgttaagcct ggaagtagag agttcaagga attagggatg
      541 taataagcgg ccgctgtatc catatgacca tgattacgaa ttcgagctcg gtacccgggg
      601 atcctctaga gtcgacgtca cgcgtccatg gagatctcga ggcctgcaga catgcaagct
      661 tggcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
      721 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
      781 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgctagcaa acttaattat
      841 ttatagtgtt acttaaaaaa tgtaaatttt ttagagtaaa aatttgtaat ataacatata
      901 acttaaaaat aaaaagcagt tagccttata aatgaaggtt agctgttttt tttgttaaaa
      961 atatatttac aaaataagag atagtagcag tacaatataa gtataaggta aaagtattat
     1021 atattaggga ggggtaagta tgaaaaatca attaaatgaa aataataaaa aggttgtatt
     1081 ttttggagta ggtgctgtag gagctacttt tgcagaacaa ttttttaact ctaaatatga
     1141 ttttaaaatt ctttgtgata atgaaagaaa gaaaagatat ttagaagaag gatttataat
     1201 aaatggaaaa aggtatgatt ttgattatgt aactaaagat gagtataaac aagaggctga
     1261 ttttataatt ataggtctaa aatataataa tttaaaagaa aatataaaag aattagatgg
     1321 attagttgga aaaaatacag ttataatgtc tctgctaaat ggagttgata gcgaagagat
     1381 aataggagaa agatttggaa ttgaaaaaat ggtatattca tatgttacca atatagatgc
     1441 aaagaaaatc aataataata ttatacacac tactaatggg ataattgtat ttggtaacaa
     1501 agataatagt gaagatagaa aaactaatat aataaccgaa gtctttgatg atgtaaatat
     1561 agaatatact ttatcaaaag atattcaacg agatatgtgg tggaagtaca tggttaatat
     1621 tggtgtaaat caaacttcag ctatacttgg tgcaccttat ggagtttttc agagttctga
     1681 gcatttaaga gaattagcaa aatctgcaat gagggaagtt gttgctatag cacaagcaaa
     1741 agacatatct cttacagaag atgatgtgga acattcatta catagaatac tagaacattc
     1801 aaaagaagga agaacatcaa tgcttcaaga tgtggaagct catagactta cagaagtaga
     1861 tatgttttct aagaatatct gtaaacttgg aaaaaaatat aacataccta ctcctataaa
     1921 tcagactttc ttttatatga taaaagtaat tgaaagtaga ttttaaaggg gctttgtaag
     1981 aaatattgat aattattgat tttaaatttt attaatgtta tatactatgg cgcgccgcca
     2041 ttattttttt gaacaattga caattcattt cttatttttt attaagtgat agtcaaaagg
     2101 cataacagtg ctgaatagaa agaaatttac agaaaagaaa attatagaat ttagtatgat
     2161 taattatact catttatgaa tgtttaattg aatacaaaaa aaaatacttg ttatgtattc
     2221 aattacgggt taaaatatag acaagttgaa aaatttaata aaaaaataag tcctcagctc
     2281 ttatatatta agctaccaac ttagtatata agccaaaact taaatgtgct accaacacat
     2341 caagccgtta gagaactcta tctatagcaa tatttcaaat gtaccgacat acaagagaaa
     2401 cattaactat atatattcaa tttatgagat tatcttaaca gatataaatg taaattgcaa
     2461 taagtaagat ttagaagttt atagcctttg tgtattggaa gcagtacgca aaggcttttt
     2521 tatttgataa aaattagaag tatatttatt ttttcataat taatttatga aaatgaaagg
     2581 gggtgagcaa agtgacagag gaaagcagta tcttatcaaa taacaaggta ttagcaatat
     2641 cattattgac tttagcagta aacattatga cttttatagt gcttgtagct aagtagtacg
     2701 aaagggggag ctttaaaaag ctccttggaa tacatagaat tcataaatta atttatgaaa
     2761 agaagggcgt atatgaaaac ttgtaaaaat tgcaaagagt ttattaaaga tactgaaata
     2821 tgcaaaatac attcgttgat gattcatgat aaaacagtag caacctattg cagtaaatac
     2881 aatgagtcaa gatgtttaca taaagggaaa gtccaatgta ttaattgttc aaagatgaac
     2941 cgatatggat ggtgtgccat aaaaatgaga tgttttacag aggaagaaca gaaaaaagaa
     3001 cgtacatgca ttaaatatta tgcaaggagc tttaaaaaag ctcatgtaaa gaagagtaaa
     3061 aagaaaaaat aatttattta ttaatttaat attgagagtg ccgacacagt atgcactaaa
     3121 aaatatatct gtggtgtagt gagccgatac aaaaggatag tcactcgcat tttcataata
     3181 catcttatgt tatgattatg tgtcggtggg acttcacgac gaaaacccac aataaaaaaa
     3241 gagttcgggg tagggttaag catagttgag gcaactaaac aatcaagcta ggatatgcag
     3301 tagcagaccg taaggtcgtt gtttaggtgt gttgtaatac atacgctatt aagatgtaaa
     3361 aatacggata ccaatgaagg gaaaagtata atttttggat gtagtttgtt tgttcatcta
     3421 tgggcaaact acgtccaaag ccgtttccaa atctgctaaa aagtatatcc tttctaaaat
     3481 caaagtcaag tatgaaatca taaataaagt ttaattttga agttattatg atattatgtt
     3541 tttctattaa aataaattaa gtatatagaa tagtttaata atagtatata cttaatgtga
     3601 taagtgtctg acagtgtcac agaaaggatg attgttatgg attataagcg gccggccagt
     3661 gggcaagttg aaaaattcac aaaaatgtgg tataatatct ttgttcatta gagcgataaa
     3721 cttgaatttg agagggaact tagatggtat ttgaaaaaat tgataaaaat agttggaaca
     3781 gaaaagagta ttttgaccac tactttgcaa gtgtaccttg tacctacagc atgaccgtta
     3841 aagtggatat cacacaaata aaggaaaagg gaatgaaact atatcctgca atgctttatt
     3901 atattgcaat gattgtaaac cgccattcag agtttaggac ggcaatcaat caagatggtg
     3961 aattggggat atatgatgag atgataccaa gctatacaat atttcacaat gatactgaaa
     4021 cattttccag cctttggact gagtgtaagt ctgactttaa atcattttta gcagattatg
     4081 aaagtgatac gcaacggtat ggaaacaatc atagaatgga aggaaagcca aatgctccgg
     4141 aaaacatttt taatgtatct atgataccgt ggtcaacctt cgatggcttt aatctgaatt
     4201 tgcagaaagg atatgattat ttgattccta tttttactat ggggaaatat tataaagaag
     4261 ataacaaaat tatacttcct ttggcaattc aagttcatca cgcagtatgt gacggatttc
     4321 acatttgccg ttttgtaaac gaattgcagg aattgataaa tagttaactt caggtttgtc
     4381 tgtaactaaa aacaagtatt taagcaaaaa catcgtagaa atacggtgtt ttttgttacc
     4441 ctaagtttaa actccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     4501 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4561 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4621 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4681 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     4741 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     4801 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     4861 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     4921 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     4981 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     5041 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     5101 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     5161 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     5221 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     5281 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcagg gccccctgct
     5341 tcggggtcat tatagcgatt ttttcggtat atccatcctt tttcgcacga tatacaggat
     5401 tttgccaaag ggttcgtgta gactttcctt ggtgtatcca acggcgtcag ccgggcagga
     5461 taggtgaagt aggcccaccc gcgagcgggt gttccttctt cactgtccct tattcgcacc
     5521 tggcggtgct caacgggaat cctgctctgc gaggctggcc ggctaccgcc ggcgtaacag
     5581 atgagggcaa gcggatggct gatgaaacca agccaaccag gaagggcagc ccacctatca
     5641 aggtgtactg ccttccagac gaacgaagag cgattgagga aaaggcggcg gcggccggca
     5701 tgagcctgtc ggcctacctg ctggccgtcg gccagggcta caaaatcacg ggcgtcgtgg
     5761 actatgagca cgtccgcgag ctggcccgca tcaatggcga cctgggccgc ctgggcggcc
     5821 tgctgaaact ctggctcacc gacgacccgc gcacggcgcg gttcggtgat gccacgatcc
     5881 tcgccctgct ggcgaagatc gaagagaagc aggacgagct tggcaaggtc atgatgggcg
     5941 tggtccgccc gagggcagag ccatgacttt tttagccgct aaaacggccg gggggtgcgc
     6001 gtgattgcca agcacgtccc catgcgctcc atcaagaaga gcgacttcgc ggagctggtg
     6061 aagtacatca ccgacgagca aggcaagacc gatcgggccc
//
